Relevant bibliographies by topics / Fluorescent screens

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  1. Journal articles
  2. Dissertations / Theses
  3. Books
  4. Book chapters
  5. Conference papers
  6. Reports

Academic literature on the topic 'Fluorescent screens'

Author: Grafiati
Published: 4 June 2021
Last updated: 28 January 2023

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Journal articles on the topic "Fluorescent screens"

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You, Kyusuk, Heping Zhu, and John Paul Abbott. "Assessment of Fluorescent Dye Brilliant Sulfaflavine Deposition on Stainless Steel Screens as Spray Droplet Collectors." Transactions of the ASABE 62, no. 2 (2019): 495–503. http://dx.doi.org/10.13031/trans.13136.

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Abstract. The fluorescent dye Brilliant Sulfaflavine (BSF, CAS 2391-30-2) was investigated to determine its photo-stability and recovery on five spray deposition collectors: white plastic plate, nylon screen, and stainless steel (SS) screens of three mesh sizes (40, 60, and 80). The photo-stability of the deposited dye was determined by measuring the variance in fluorescence intensity after daylight exposure. The recovery rates were investigated with statically dispensed droplets and dynamically discharged droplets. In addition, droplet penetration through the screen collectors and the amount of unrecovered dye on reprocessed collectors were assessed to better understand the differences in dye recovery rates among different collector types. Photo-degradation tests verified that all collector types were insignificant in fluorescence degradation (<3.1%) after 120 min of solar exposure. Nylon screens had the lowest dye recovery rate (87.0%) for statically dispensed droplets, whereas plastic plates and SS screens recovered more than 90% of deposited dye. For dynamically discharged droplets, the 60-mesh and 80-mesh SS screens recovered more than 70% of the deposited dye, whereas nylon screens showed less than 50% recovery rate. These results were substantiated visually with a high-speed imaging system that detected droplets penetrating more frequently through screens with larger mesh openings. Throughout ten continuous reprocessing cycles of the fluorimetry test, the fluorescence intensity on reprocessed nylon and 80-mesh SS screen collectors increased by 16.8% and 12.7%, respectively, while there was less than 1% change in fluorescence intensity on the 40-mesh and 60-mesh SS screens. These results were clarified through dye residue verification using digital image analysis. Keywords: Fluorescence intensity, Photo-degradation, Recovery rate, Spray deposition assessment, Stainless steel screen.
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Schaaf, Tory M., Evan Kleinboehl, Samantha L. Yuen, Lauren N. Roelike, Bengt Svensson, Andrew R. Thompson, Razvan L. Cornea, and David D. Thomas. "Live-Cell Cardiac-Specific High-Throughput Screening Platform for Drug-Like Molecules That Enhance Ca2+ Transport." Cells 9, no. 5 (May 8, 2020): 1170. http://dx.doi.org/10.3390/cells9051170.

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We engineered a concatenated fluorescent biosensor and dual-wavelength fluorescence lifetime (FLT) detection, to perform high-throughput screening (HTS) in living cells for discovery of potential heart-failure drugs. Heart failure is correlated with insufficient activity of the sarcoplasmic reticulum Ca-pump (SERCA2a), often due to excessive inhibition by phospholamban (PLB), a small transmembrane protein. We sought to discover small molecules that restore SERCA2a activity by disrupting this inhibitory interaction between PLB and SERCA2a. Our approach was to fluorescently tag the two proteins and measure fluorescence resonance energy transfer (FRET) to detect changes in binding or structure of the complex. To optimize sensitivity to these changes, we engineered a biosensor that concatenates the two fluorescently labeled proteins on a single polypeptide chain. This SERCA2a-PLB FRET biosensor construct is functionally active and effective for HTS. By implementing 2-wavelength FLT detection at extremely high speed during primary HTS, we culled fluorescent compounds as false-positive Hits. In pilot screens, we identified Hits that alter the SERCA2a-PLB interaction, and a newly developed secondary calcium uptake assay revealed both activators and inhibitors of Ca-transport. We are implementing this approach for large-scale screens to discover new drug-like modulators of SERCA2a-PLB interactions for heart failure therapeutic development.
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Ljosa, Vebjorn, and Anne E. Carpenter. "High-throughput screens for fluorescent dye discovery." Trends in Biotechnology 26, no. 10 (October 2008): 527–30. http://dx.doi.org/10.1016/j.tibtech.2008.06.008.

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Horvath, Peter, Thomas Wild, Ulrike Kutay, and Gabor Csucs. "Machine Learning Improves the Precision and Robustness of High-Content Screens." Journal of Biomolecular Screening 16, no. 9 (August 1, 2011): 1059–67. http://dx.doi.org/10.1177/1087057111414878.

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Imaging-based high-content screens often rely on single cell-based evaluation of phenotypes in large data sets of microscopic images. Traditionally, these screens are analyzed by extracting a few image-related parameters and use their ratios (linear single or multiparametric separation) to classify the cells into various phenotypic classes. In this study, the authors show how machine learning–based classification of individual cells outperforms those classical ratio-based techniques. Using fluorescent intensity and morphological and texture features, they evaluated how the performance of data analysis increases with increasing feature numbers. Their findings are based on a case study involving an siRNA screen monitoring nucleoplasmic and nucleolar accumulation of a fluorescently tagged reporter protein. For the analysis, they developed a complete analysis workflow incorporating image segmentation, feature extraction, cell classification, hit detection, and visualization of the results. For the classification task, the authors have established a new graphical framework, the Advanced Cell Classifier, which provides a very accurate high-content screen analysis with minimal user interaction, offering access to a variety of advanced machine learning methods.
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Kolb, Janet M., Gregory Yamanaka, and Susan P. Manly. "Use of a Novel Homogeneous Fluorescent Technology in High Throughput Screening." Journal of Biomolecular Screening 1, no. 4 (June 1996): 203–10. http://dx.doi.org/10.1177/108705719600100407.

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A new fluorescent technology called homogeneous time-resolved fluorescence (HTRF) is sensitive, homogeneous, and quite tolerant to extremes in reaction conditions. These characteristics make this technique an attractive candidate for use in high throughput screens. The assay system uses a pair of fluorescent compounds to label biomolecules. The long-lived nature of the fluorescence of one of them, europium cryptate, facilitates the homogeneous nature of the assay. Furthermore, the introduction of a time delay in reading the signal eliminates the principal difficulty in applying fluorescence to screening formats, that of endogenous fluorescence of samples tested (especially natural products). This technique is robust and sensitive, and we report here its utility in a high throughput screening format.
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George, Jeanette, Michelle L. Teear, Christopher G. Norey, and D. Dougal Burns. "Evaluation of an Imaging Platform during the Development of a FRET Protease Assay." Journal of Biomolecular Screening 8, no. 1 (January 2003): 72–80. http://dx.doi.org/10.1177/1087057102239778.

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Synthetic peptide substrates labeled with a fluorescent donor and quenching moiety flanking an enzyme cleavage site provide a reliable method for monitoring enzyme activity. The dye pair Mca/Dnp has been widely used for this purpose, but poor solubility characteristics, combined with fluorescence emission in the region of the spectrum associated with interference from bi-ologicals and library compounds, can limit the usefulness of Mca/Dnp substrates in a high-throughput screening (HTS) environment. Peptide Mca-Arg-Pro-Lys-Pro-Val-Glu-Nva-Trp-Arg-Lys(Dnp)-NH2 is a matrix-metalloproteinase 3 (MMP-3) enzyme substrate that the authors have labeled with a CyDye pair, Cy3/Cy5Q. The Mca/Dnp- and CyDye-labeled substrates were compared during the development of an MMP-3 inhibitor assay. The results obtained showed that although the peptide substrates behaved similarly throughout the development of the MMP-3 assay, during a test screen of 934 compounds randomly selected from a collection of more than 70,000 compounds, the CyDye substrate was considerably more reliable. Screen Z factor values of 0.84 and 0.15 were obtained using the CyDye and Mca/Dnp peptides respectively, and the authors found that although < 1% of the test compounds were auto-fluorescent at Cy3 wavelengths, > 10% could not be screened using the Mca/Dnp substrate because of compound auto-fluorescence and interference. During this study, the authors used a PMTbased fluorescence plate reader and at the same time evaluated a charged couple device (CCD)—based imaging platform specifically optimized for use with CyDye reagents. The imaging platform gave improved read accuracy and faster plate processing times compared with the PMT reader. Overall, the results presented here highlight the potential benefit of employing the red-shifted CyDye reagents and imaging technology during the development and execution of HTS protease screens. (Journal of Biomolecular Screening 2003:72-80)
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Scott, Benjamin M., Leanne E. Wybenga-Groot, C. Jane McGlade, Elise Heon, Sergio G. Peisajovich, and Belinda S. W. Chang. "Screening of Chemical Libraries Using a Yeast Model of Retinal Disease." SLAS DISCOVERY: Advancing the Science of Drug Discovery 24, no. 10 (September 26, 2019): 969–77. http://dx.doi.org/10.1177/2472555219875934.

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Retinitis pigmentosa (RP) is a degenerative retinal disease, often caused by mutations in the G-protein-coupled receptor rhodopsin. The majority of pathogenic rhodopsin mutations cause rhodopsin to misfold, including P23H, disrupting its crucial ability to respond to light. Previous screens to discover pharmacological chaperones of rhodopsin have primarily been based on rescuing rhodopsin trafficking and localization to the plasma membrane. Here, we present methods utilizing a yeast-based assay to screen for compounds that rescue the ability of rhodopsin to activate an associated downstream G-protein signaling cascade. We engineered a yeast strain in which human rhodopsin variants were genomically integrated, and were able to demonstrate functional coupling to the yeast mating pathway, leading to fluorescent protein expression. We confirmed that a known pharmacological chaperone, 9- cis retinal, could partially rescue light-dependent activation of a disease-associated rhodopsin mutation (P23H) expressed in yeast. These novel yeast strains were used to perform a phenotypic screen of 4280 compounds from the LOPAC1280 library and a peptidomimetic library, to discover novel pharmacological chaperones of rhodopsin. The fluorescence-based assay was robust in a 96-well format, with a Z′ factor of 0.65 and a signal-to-background ratio of above 14. One compound was selected for additional analysis, but it did not appear to rescue rhodopsin function in yeast. The methods presented here are amenable to future screens of small-molecule libraries, as they are robust and cost-effective. We also discuss how these methods could be further modified or adapted to perform screens of more compounds in the future.
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Giakoumakis, G. E., and D. M. Miliotis. "Light angular distribution of fluorescent screens excited by x-rays." Physics in Medicine and Biology 30, no. 1 (January 1, 1985): 21–29. http://dx.doi.org/10.1088/0031-9155/30/1/003.

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Talbot, Jan B. "From the Editor: Let There Be Light." Electrochemical Society Interface 7, no. 2 (June 1, 1998): 3. http://dx.doi.org/10.1149/2.001982if.

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Certain materials can absorb energy from such sources as X-rays, ultraviolet light, or electrons, then re-emit the energy as visible light. These luminescent materials, called “phosphors,” are ubiquitous. Phosphors light up fluorescent light bulbs, television screens, computer screens, and signs and displays everywhere we look. These are the materials of interest to the Luminescence and Display Materials Division, featured in this issue.
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FANELLI, ANTHONY J., JOAN V. BURLEW, and MINA K. GABRIEL. "Protection of Milk Packaged in High Density Polyethylene Against Photodegradation by Fluorescent Light." Journal of Food Protection 48, no. 2 (February 1, 1985): 112–17. http://dx.doi.org/10.4315/0362-028x-48.2.112.

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The effectiveness of visible and UV light screens, compounded in polyethylene dairy resin to protect vitamins in milk from photodegradation, was investigated. Three pigments and three UV absorbers were chosen for testing on the basis of their commercial availability, FDA approval for contact with food, and advertised compatibility with polyolefins. In this study, vitamin decomposition was accelerated over what would be experienced in a commercial milk container in order to expedite the testing program and exaggerate differences in effectiveness of the various light screens. Good protection of vitamin A and riboflavin was provided by 0.3 wt % FD&C yellow #5. Protection of ascorbic acid was marginal. Two of the UV absorbers, Cyasorb 531 and Tinuvin 326, afforded protection of vitamin A, but not riboflavin or ascorbic acid. Visible and UV spectra are presented for the vitamins and light screens used in this work.
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Dissertations / Theses on the topic "Fluorescent screens"

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Cardno, Tony Stuart, and n/a. "Development of a high throughput fluorescent screening assay for genetic recoding." University of Otago. Department of Biochemistry, 2007. http://adt.otago.ac.nz./public/adt-NZDU20071218.145806.

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The development of new drug therapies traditionally requires mass screening of thousands if not millions of substances to identify lead compounds. They are then further optimised to increase potency. The screening of the large pharmaceutical compound libraries can be incredibly expensive, with the industry responding by miniaturising the assays to smaller formats, enabling the compound screening to be automated and, importantly, eliminating assay reagents that are a major contributing cost for running large screens. A potential target for such an approach is the genetic recoding site of viruses like HIV-1 and SARS. They use programmed recoding of the genetic code to regulate the translation of necessary proteins required for viable virus production. For example HIV-1 uses a -1 frameshift mechanism to regulate the ratio of the Gag to the Pol proteins, crucial for viable virus formation. The study of recoding, including readthrough of premature termination codons have most recently used bicistronic reporters with different combinations of enzymes. The most widely used plasmid bicistronic reporter utilises a dual luciferase arrangement comprised of firefly luciferase and Renilla luciferase reporters flanking the DNA being studied. Both of the luciferase enzymatic reporters emit light in response to their respective substrates. The cost of these substrates is the major issue to using luciferase reporters for high throughput screening. My study aimed at designing and developing a bicistronic assay suitable for genetic recoding that was amenable to high throughput screening. The luciferase reporters were replaced with Green Fluorescent Protein (GFP) reporters that do not require the addition of substrates. The development of a dual GFP assay required the appropriate selection of GFP fluorophores, the best arrangement of the GFPs to maximise the ratio of relative fluorescence intensity signal to background, the optimisation of the cells and growth conditions, DNA transfection, plate reader selection, and optical filter sets. Cassettes encoding protein linkers were also incorporated into the design of the constructs to separate the fluorescent proteins spatially to facilitate unimpaired folding into their functional units within the fusion protein. The assay was further improved by moving from transient transfection to stably expressing cell lines. A viable assay was almost achieved for 96 (and 384) well plates with a Z� factor compatible with the assay being suitable for high throughput screening. The assay was used to test a small collection of compounds known to interact with the ribosome and compounds known in the literature to affect frameshifting. This proof of concept was important, since it showed that the assay, with the various modifications, optimisations and miniaturisation steps, still retained the capability of correctly measuring the -1 frameshifting efficiency at the HIV-1 recoding site, and recording compound-induced modulations to the frameshifting efficiency. The compounds cycloheximide and anisomycin, for example, were shown to decrease -1 frameshifting albeit at some expense to overall protein synthesis. The dual GFP assay was also shown to be able to measure accurately changes in the frameshift efficiency brought about by mutations to the frameshift element, and additionally, it would be suitable for the detection and study of compounds, like the recently reported PTC-124 (currently undergoing phase II clinical trial for Duchenne Muscular Dystrophy and cystic fibrosis) that increases readthrough of a UGA premature stop codon mutation. The dual GFP assay developed in this study is at most only 1/10th of the cost of a comparable dual luciferase assay, largely due to removal of assay substrates and transfection reagents. The assay has a robust Z� factor comparable to that of the dual luciferase assay, and would substantially decrease the costs of high throughput screening in situations where a bicistronic reporter is required. The HIV-1 frameshift element is such a site.
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Perry, Kyle James. "Differential fluorescence-based genetic screens to identify novel Listeria monocytogenes virulence determinants." Thesis, Harvard University, 2015. http://nrs.harvard.edu/urn-3:HUL.InstRepos:14226063.

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Listeria monocytogenes is a Gram-positive, facultative intracellular pathogen that causes gastroenteritis, which in the young, the elderly, and the immunocompromised can progress to severe invasive disease with high mortality. While previous studies have largely elucidated the bacterial and host mechanisms necessary for the bacterium to access its replicative niche in the host cell cytosol, the L. monocytogenes factors required for adaptation to life within this restrictive environment are poorly understood. In this dissertation, I describe a fluorescence-activated cell sorting (FACS)-based differential fluorescence genetic screening technique for the identification of L. monocytogenes genes necessary for optimal intracellular replication. Bacteria harboring deletions in identified genes were defective for intracellular replication, plaque formation, and in vivo virulence, validating the ability of the screening method to identify novel intracellular replication-defective mutants. Minor alteration of the FACS-based screening strategy allowed the detection and differentiation of bacterial mutants displaying varying severities of actin-based motility defects. A preliminary FACS-based genetic screen to identify actin-based motility mutants isolated multiple independent insertions within internal control genes, demonstrating the potential utility of FACS-based differential fluorescence genetic screening methods for the identification of L. monocytogenes genes important for multiple virulence phenotypes. Lastly, my characterization of the X-prolyl aminopeptidase PepP, a novel virulence factor identified by the FACS-based genetic screen to discover genes necessary for optimal intracellular replication, revealed this enzyme plays an unexpected role in L. monocytogenes virulence gene regulation.
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Star, Eleanor. "Use of VEGF splicing-sensitive fluorescent reporters to screen for anti-angiogenic molecules." Thesis, University of Bristol, 2016. http://hdl.handle.net/1983/ba789d02-7085-4423-9489-362f147c9dec.

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Alternative splicing of the vascular endothelial growth factor A (VEGF-A) terminal exon generates two protein families with differing functions. Pro-angiogenic VEGFxxx isoforms are produced via selection of the proximal 3’ splice site of the terminal exon. Use of an alternative distal splice site creates the anti-angiogenic VEGFxxxb proteins. A bichromatic splicing-sensitive reporter was designed to mimic VEGF alternative splicing and used as a molecular tool to further investigate this alternative splicing event. The terminal exon and preceding intron of VEGF were inserted into a minigene construct followed by the coding sequences for two fluorescent proteins. A different fluorescent protein is expressed depending on which 3’ splice site of the exon is used during splicing. The fluorescent output can be used to follow splicing decisions in vitro and in vivo. Following successful reporter validation in different cell lines and altering splicing using known modulators, small pilot screens were undertaken to search for novel regulators of the splicing decision that creates pro-/anti-angiogenic VEGF isoforms. A larger screen was performed using a library of 1280 small molecules (LOPAC), all compounds of the library were pharmacologically active and have known biological targets. Alterations to reporter splicing were measured using a fluorescent plate reader to detect RFP and GFP expression. Compounds of interest were further validated using flow cytometry and assessed for effect on endogenous VEGF alternative splicing. In vitro angiogenesis assays were used to demonstrate anti-angiogenic effect. Anti-angiogenic activity and the effect on tumour growth were investigated in several in vivo models.
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Gorzitze-Maxey, Adrian Dale. "Development of a Fluorescence-Based Screen for Glutathione-S-Transferase Inhibitors." Ohio Dominican University Honors Theses / OhioLINK, 2015. http://rave.ohiolink.edu/etdc/view?acc_num=oduhonors1449612834.

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Martin, Sarah Friede. "Fluorescence resonance energy transfer studies of protein interactions." Thesis, St Andrews, 2008. http://hdl.handle.net/10023/537.

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Wang, You. "Development of yeast-based methods to screen for plant cytokinin-binding proteins." Access electronically, 2004. http://www.library.uow.edu.au/adt-NWU/public/adt-NWU20060123.141512/index.html.

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Bakker, Jimmy W. P. "Optical Detection Using Computer Screen Photo-assisted Techniques and Ellipsometry." Doctoral thesis, Linköpings universitet, Tillämpad optik, 2006. http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-6392.

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Two main subjects, ellipsometry and computer screen photo-assisted techniques (CSPT), form the main line in this thesis. Ellipsometry is an optical technique based on the detection of polarization changes of light upon interaction with a sample. As most optical detection techniques it is non-intrusive and an additional advantage is its high surface sensitivity: thickness resolution in the order of pm can in principle be achieved. Therefore, ellipsometry is widely used as a technique for determination of optical constants and layer thickness for thin-layer structures. Lately ellipsometry has also been proposed for sensing applications, utilizing the detection of changes in the properties of thin layers. One application is described in this thesis concerning the detection of volatile organic solvents in gas phase using modified porous silicon layers, fabricated by electrochemical etching of silicon to create nm-sized pores. This greatly increases the surface area, promoting gas detection because the number of adsorption sites increases. Other applications of ellipsometry discussed in this thesis are based on combination with CSPT. CSPT is a way to exploit existing optical techniques for use in low-cost applications. In CSPT the computer screen itself is used as a (programmable) light source for optical measurements. For detection a web camera can be used and the whole measurement platform is formed by the computer. Since computers are available almost everywhere, this is a promising way to create optical measurement techniques for widespread use, for example in home-diagnostics. Since the only thing that needs to be added is a sample holder governing the physical or chemical process and directing the light, the cost can be kept very low. First, the use of CSPT for the measurement of fluorescence is described. Fluorescence is used in many detection applications, usually by chemically attaching a fluorescent marker molecule to a suitable species in the process and monitoring the fluorescent emission. The detection of fluorescence is shown to be possible using CSPT, first in a cuvette-based setup, then using a custom designed micro array. In the latter, polarizers were used for contrast enhancement, which in turn led to the implementation of an existing idea to test CSPT for ellipsometry measurements. In a first demonstration, involving thickness measurement of silicon dioxide on silicon, a thickness resolution in the order of nm was already achieved. After improvement of the system, gradients in protein layers could be detected, opening the door toward biosensor applications. Some further development will be needed to make the CSPT applications described here ready for the market, but the results so far are certainly promising.
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Gupta, Nilay. "Localization Study of Supervillin in Zebrafish Hair Cells Using Immuno-fluorescence Assay & Identification of Small Molecules that Impact the Innervation of the Lateral Line System of Developing Zebrafish." Case Western Reserve University School of Graduate Studies / OhioLINK, 2016. http://rave.ohiolink.edu/etdc/view?acc_num=case1459898664.

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Polcer, Simon. "Detekce a rozměření elektronového svazku v obrazech z TEM." Master's thesis, Vysoké učení technické v Brně. Fakulta elektrotechniky a komunikačních technologií, 2020. http://www.nusl.cz/ntk/nusl-413022.

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This diploma thesis deals with automatic detection and measurement of the electron beam in the images from a transmission electron microscope (TEM). The introduction provides a description of the construction and the main parts of the electron microscope. In the theoretical part, there are summarized modes of illumination from the fluorescent screen. Machine learning, specifically convolution neural network U-Net is used for automatic detection of the electron beam in the image. The measurement of the beam is based on ellipse approximation, which defines the size and dimension of the beam. Neural network learning requires an extensive database of images. For this purpose, the own augmentation approach is proposed, which applies a specific combination of geometric transformations for each mode of illumination. In the conclusion of this thesis, the results are evaluated and summarized. This proposed algorithm achieves 0.815 of the DICE coefficient, which describes an overlap between two sets. The thesis was designed in Python programming language.
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Hodgkinson, Mark. "Cause and control of oil induced phytotoxicity." Thesis, Queensland University of Technology, 1999.

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Books on the topic "Fluorescent screens"

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Cathodoluminescence and photoluminescence: Theories and practical applications. Boca Raton, FL: CRC Press, 2007.

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Ivanov, A. P. Optika li͡u︡minest͡s︡entnogo ėkrana. Minsk: Nauka i tekhnika, 1985.

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Porter, Louis G. Final evaluation of a color calibrator for a radar remote weather display system / Louis G. Porter ; sponsored by Federal Aviation Administration, U.S. Department of Transportation. [Washington, D.C.]: U.S. Dept. of Commerce, National Bureau of Standards, 1986.

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Lu se ying ping: Zhongguo nong ye ying shi er shi nian chuang zuo ji = Green fluorescent screen : records of Chinese agricultural film & TV creations in 20 years. Beijing Shi: Zhongguo chuan mei da xue chu ban she, 2005.

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Ozawa, Lyuji. Cathodoluminescence and Photoluminescence: Theories and Practical Applications. Taylor & Francis Group, 2018.

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Ozawa, Lyuji. Cathodoluminescence and Photoluminescence: Theories and Practical Applications (Phosphor Science and Engineering). CRC, 2007.

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Ozawa, Lyuji. Cathodoluminescence and Photoluminescence. Taylor & Francis Group, 2019.

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Ozawa, Lyuji. Cathodoluminescence and Photoluminescence: Theories and Practical Applications. Taylor & Francis Group, 2018.

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Ozawa, Lyuji. Cathodoluminescence and Photoluminescence: Theories and Practical Applications. Taylor & Francis Group, 2018.

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Ozawa, Lyuji. Cathodoluminescence and Photoluminescence: Theories and Practical Applications. Taylor & Francis Group, 2018.

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Book chapters on the topic "Fluorescent screens"

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Josan, Jatinder S., Channa R. De Silva, Byunghee Yoo, Ronald M. Lynch, Mark D. Pagel, Josef Vagner, and Victor J. Hruby. "Fluorescent and Lanthanide Labeling for Ligand Screens, Assays, and Imaging." In Methods in Molecular Biology, 89–126. Totowa, NJ: Humana Press, 2011. http://dx.doi.org/10.1007/978-1-61779-012-6_6.

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Warzecha, Claude C., Ruben Hovhannisyan, and Russ P. Carstens. "Dynamic Fluorescent and Luminescent Reporters for Cell-Based Splicing Screens." In Methods in Molecular Biology, 273–87. Totowa, NJ: Humana Press, 2012. http://dx.doi.org/10.1007/978-1-61779-767-5_18.

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Briani, Federica. "Cell-Based Fluorescent Screen to Identify Inhibitors of Bacterial Translation Initiation." In Methods in Molecular Biology, 237–45. New York, NY: Springer New York, 2016. http://dx.doi.org/10.1007/978-1-4939-6634-9_14.

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Sanchez, Lisa, Yuen-Yan Chang, Nora Mellouk, and Jost Enninga. "Time-Resolved Fluorescence Microscopy Screens on Host Protein Subversion During Bacterial Cell Invasion." In Methods in Molecular Biology, 113–31. New York, NY: Springer US, 2022. http://dx.doi.org/10.1007/978-1-0716-2449-4_8.

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Raneri, Matteo, Emilio Alvarez-Ruiz, Danuta Mossakovska, and Federica Briani. "Cell-Based Fluorescent Screen Amenable to HTS to Identify Inhibitors of Bacterial Translation Initiation." In Methods in Molecular Biology, 303–12. New York, NY: Springer US, 2022. http://dx.doi.org/10.1007/978-1-0716-2855-3_16.

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Umetani, Keiji, Tohoru Takeda, Hironori Ueki, Ken Ueda, Yuji Itai, Izumi Anno, Teiichi Nakajima, and Masayoshi Akisada. "Iodine-Filter Imaging System for Subtraction Angiography and Its Improvement by Fluorescent-Screen Harpicon Detector." In Medical Applications of Synchrotron Radiation, 99–102. Tokyo: Springer Japan, 1998. http://dx.doi.org/10.1007/978-4-431-68485-5_16.

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Humbert, Nicolas, and Thomas R. Ward. "Functionality Screen of Streptavidin Mutants by Non-Denaturing SDS–PAGE Using Biotin-4-Fluorescein." In Avidin-Biotin Interactions, 63–71. Totowa, NJ: Humana Press, 2008. http://dx.doi.org/10.1007/978-1-59745-579-4_6.

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Miranda, Tina B., Ty C. Voss, and Gordon L. Hager. "High-Throughput Fluorescence-Based Screen to Identify Factors Involved in Nuclear Receptor Recruitment to Response Elements." In Imaging Gene Expression, 3–12. Totowa, NJ: Humana Press, 2013. http://dx.doi.org/10.1007/978-1-62703-526-2_1.

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Tchelet, Daniel, and Dor Salomon. "A Rapid Fluorescence-Based Screen to Identify Regulators and Components of Interbacterial Competition Mechanisms in Bacteria." In Methods in Molecular Biology, 11–24. New York, NY: Springer US, 2022. http://dx.doi.org/10.1007/978-1-0716-1971-1_2.

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Comyn, Sophie A., and Thibault Mayor. "A Method to Monitor Protein Turnover by Flow Cytometry and to Screen for Factors that Control Degradation by Fluorescence-Activated Cell Sorting." In Methods in Molecular Biology, 137–53. New York, NY: Springer New York, 2018. http://dx.doi.org/10.1007/978-1-4939-8706-1_10.

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Conference papers on the topic "Fluorescent screens"

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Silva, Tiago F., Cristiane Jahnke, Zwinglio O. Guimarães, Marcos N. Martins, and Vito R. Vanin. "Saturation effects on beam diagnostics using fluorescent screens." In XXXIII BRAZILIAN WORKSHOP ON NUCLEAR PHYSICS. AIP, 2011. http://dx.doi.org/10.1063/1.3608965.

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Hillen, Walter, W. Eckenbach, Peter Quadflieg, and Thomas T. Zaengel. "Signal-to-noise performance in cesium iodide x-ray fluorescent screens." In Medical Imaging '91, San Jose, CA, edited by Roger H. Schneider. SPIE, 1991. http://dx.doi.org/10.1117/12.43435.

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Berg, W., and K. Ko. "Fluorescent screens and image processing for the APS linac test stand." In Accelerator instrumentation fourth annual workshop. AIP, 1992. http://dx.doi.org/10.1063/1.44348.

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Berg, W., and K. Ko. "Status of the fluorescent screens and image processing for the APS Linac." In Beam Instrumentation Workshop. AIP, 1994. http://dx.doi.org/10.1063/1.46982.

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You, Kyusuk, Heping Zhu, and JohnPaul R. Abbott. "Recovery rates of fluorescent dye on screens and plates as spray deposition collectors." In 2018 Detroit, Michigan July 29 - August 1, 2018. St. Joseph, MI: American Society of Agricultural and Biological Engineers, 2018. http://dx.doi.org/10.13031/aim.201800066.

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Rakkolainen, Ismo, and Karri Palovuori. "A Fluorescent Mid-air Screen." In 2013 IEEE International Symposium on Multimedia (ISM). IEEE, 2013. http://dx.doi.org/10.1109/ism.2013.14.

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Satoh, Noritaka, Masahiro Kusakabe, Katsutoshi Ohno, Nozomu Kamagata, and Masahiro Endo. "High-luminance fluorescent screen with interference filter." In Medical Imaging 1995, edited by Richard L. Van Metter and Jacob Beutel. SPIE, 1995. http://dx.doi.org/10.1117/12.208366.

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Park, Sanghan, Satya Gowthami Achanta, and Chang-Soo Kim. "Fluorescence intensity measurements with display screen as excitation source." In SPIE Defense, Security, and Sensing, edited by Brian M. Cullum and Eric S. McLamore. SPIE, 2011. http://dx.doi.org/10.1117/12.885175.

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Qiu, YaFeng, BenKang Chang, YunSheng Qian, RongGuo Fu, Youtang Gao, and Tian Si. "Model for the brightness uniformity of fluorescence screen of image intensifier." In 27th International congress on High-Speed Photography and Photonics, edited by Xun Hou, Wei Zhao, and Baoli Yao. SPIE, 2007. http://dx.doi.org/10.1117/12.725237.

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Kumagai, Kota, Satoshi Hasegawa, and Yoshio Hayasaki. "Volumetric display with holographic parallel two-photon excitations and multilayer fluorescence screen." In Digital Holography and Three-Dimensional Imaging. Washington, D.C.: OSA, 2015. http://dx.doi.org/10.1364/dh.2015.dw3a.2.

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Reports on the topic "Fluorescent screens"

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Johnson, C. D. Limits to the resolution of beam size measurement from fluorescent screens due to the thickness of the phosphor. Office of Scientific and Technical Information (OSTI), July 1988. http://dx.doi.org/10.2172/6776306.

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Sadot, Einat, Christopher Staiger, and Zvi Kam Weizmann. functional genomic screen for new plant cytoskeletal proteins and the determination of their role in actin mediated functions and guard cells regulation. United States Department of Agriculture, January 2003. http://dx.doi.org/10.32747/2003.7587725.bard.

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The original objectives of the approved proposal were: 1. To construct a YFP fused Arabidopsis cDNA library in a mammalian expression vector. 2. To infect the library into a host fibroblast cell line and to screen for new cytoskeletal associated proteins using an automated microscope. 3. Isolate the new genes. 4. Characterize their role in plants. The project was approved as a feasibility study to allow proof of concept that would entail building the YFP library and picking up a couple of positive clones using the fluorescent screen. We report here on the construction of the YFP library, the development of the automatic microscope, the establishment of the screen and the isolation of positive clones that are plant cDNAs encoding cytoskeleton associated proteins. The rational underling a screen of plant library in fibroblasts is based on the high conservation of the cytoskeleton building blocks, actin and tubulin, between the two kingdoms (80-90% homology at the level of amino acids sequence). In addition, several publications demonstrated the recognition of mammalian cytoskeleton by plant cytoskeletal binding proteins and vice versa. The major achievements described here are: 1. The development of an automated microscope equipped with fast laser auto-focusing for high magnification and a software controlling 6 dimensions; X, Y position, auto focus, time, color, and the distribution and density of the fields acquired. This system is essential for the high throughput screen. 2. The construction of an extremely competent YFP library efficiently cloned (tens of thousands of clones collected, no empty vectors detected) with all inserts oriented 5't03'. These parameters render it well representative of the whole transcriptome and efficient in "in-frame" fusion to YFP. 3. The strategy developed for the screen allowing the isolation of individual positive cDNA clones following three rounds of microscopic scans. The major conclusion accomplished from the work described here is that the concept of using mammalian host cells for fishing new plant cytoskeletal proteins is feasible and that screening system developed is complete for addressing one of the major bottlenecks of the plant cytoskeleton field: the need for high throughput identification of functionally active cytoskeletal proteins. The new identified plant cytoskeletal proteins isolated in the pilot screen and additional new proteins which will be isolated in a comprehensive screen will shed light on cytoskeletal mediated processes playing a major role in cellular activities such as cell division, morphogenesis, and functioning such as chloroplast positioning, pollen tube and root hair elongation and the movement of guard cells. Therefore, in the long run the screen described here has clear agricultural implications.
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Zchori-Fein, Einat, Judith K. Brown, and Nurit Katzir. Biocomplexity and Selective modulation of whitefly symbiotic composition. United States Department of Agriculture, June 2006. http://dx.doi.org/10.32747/2006.7591733.bard.

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Whiteflies are sap-sucking insects that harbor obligatory symbiotic bacteria to fulfill their dietary needs, as well as a facultative microbial community with diverse bacterial species. The sweetpotato whitefly Bemisia tabaci (Gennadius) is a severe agricultural pest in many parts of the world. This speciesconsists of several biotypes that have been distinguished largely on the basis of biochemical or molecular diagnostics, but whose biological significance is still unclear. The original objectives of the project were (i) to identify the specific complement of prokaryotic endosymbionts associated with select, well-studied, biologically and phylogeographically representative biotypes of B. tabaci, and (ii) to attempt to 'cure’ select biotypes of certain symbionts to permit assessment of the affect of curing on whitefly fitness, gene flow, host plant preference, and virus transmission competency.To identify the diversity of bacterial community associated with a suite of phylogeographically-diverseB. tabaci, a total of 107 populations were screened using general Bacteria primers for the 16S rRNA encoding gene in a PCR. Sequence comparisons with the available databases revealed the presence of bacteria classified in the: Proteobacteria (66%), Firmicutes (25.70%), Actinobacteria (3.7%), Chlamydiae (2.75%) and Bacteroidetes (<1%). Among previously identified bacteria, such as the primary symbiont Portiera aleyrodidarum, and the secondary symbionts Hamiltonella, Cardinium and Wolbachia, a Rickettsia sp. was detected for the first time in this insect family. The distribution, transmission, and localization of the Rickettsia were studied using PCR and fluorescence in situ hybridization (FISH). Rickettsia was found in all 20 Israeli B. tabaci populations screened as well as some populations screened in the Arizona laboratory, but not in all individuals within each population. FISH analysis of B. tabaci eggs, nymphs and adults, revealed a unique concentration of Rickettsia around the gut and follicle cells as well as its random distribution in the haemolymph, but absence from the primary symbiont housing cells, the bacteriocytes. Rickettsia vertical transmission on the one hand and its partial within-population infection on the other suggest a phenotype that is advantageous under certain conditions but may be deleterious enough to prevent fixation under others.To test for the possible involvement of Wolbachia and Cardiniumin the reproductive isolation of different B. tabacibiotypes, reciprocal crosses were preformed among populations of the Cardinium-infected, Wolbachia-infected and uninfected populations. The crosses results demonstrated that phylogeographically divergent B. tabaci are reproductively competent and that cytoplasmic incompatibility inducer-bacteria (Wolbachia and Cardinium) both interfered with, and/or rescued CI induced by one another, effectively facilitating bidirectional female offspring production in the latter scenario.This knowledge has implications to multitrophic interactions, gene flow, speciation, fitness, natural enemy interactions, and possibly, host preference and virus transmission. Although extensive and creative attempts undertaken in both laboratories to cure whiteflies of non-primary symbionts have failed, our finding of naturally uninfected individuals have permitted the establishment of Rickettsia-, Wolbachia- and Cardinium-freeB. tabaci lines, which are been employed to address various biological questions, including determining the role of these bacteria in whitefly host biology.
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Shapira, Roni, Judith Grizzle, Nachman Paster, Mark Pines, and Chamindrani Mendis-Handagama. Novel Approach to Mycotoxin Detoxification in Farm Animals Using Probiotics Added to Feed Stuffs. United States Department of Agriculture, May 2010. http://dx.doi.org/10.32747/2010.7592115.bard.

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T-2 toxin, a toxic product belongs to the trichothecene mycotoxins, attracts major interest because of its severe detrimental effects on the health of human and farm animals. The occurrence of trichothecenes contamination is global and they are very resistant to physical or chemical detoxification techniques. Trichothecenes are absorbed in the small intestine into the blood stream. The hypothesis of this project was to develop a protecting system using probiotic bacteria that will express trichothecene 3-O-acetyltransferase (Tri101) that convert T-2 to a less toxic intermediate to reduce ingested levels in-situ. The major obstacle that we had faced during the project is the absence of stable and efficient expression vectors in probiotics. Most of the project period was invested to screen and isolate strong promoter to express high amounts of the detoxify enzyme on one hand and to stabilize the expression vector on the other hand. In order to estimate the detoxification capacity of the isolated promoters we had developed two very sensitive bioassays.The first system was based on Saccharomyces cerevisiae cells expressing the green fluorescent protein (GFP). Human liver cells proliferation was used as the second bioassay system.Using both systems we were able to prove actual detoxification on living cells by probiotic bacteria expressing Tri101. The first step was the isolation of already discovered strong promoters from lactic acid bacteria, cloning them downstream the Tri101 gene and transformed vectors to E. coli, a lactic acid bacteria strain Lactococcuslactis MG1363, and a probiotic strain of Lactobacillus casei. All plasmid constructs transformed to L. casei were unstable. The promoter designated lacA found to be the most efficient in reducing T-2 from the growth media of E. coli and L. lactis. A prompter library was generated from L. casei in order to isolate authentic probiotic promoters. Seven promoters were isolated, cloned downstream Tri101, transformed to bacteria and their detoxification capability was compared. One of those prompters, designated P201 showed a relatively high efficiency in detoxification. Sequence analysis of the promoter region of P201 and another promoter, P41, revealed the consensus region recognized by the sigma factor. We further attempted to isolate an inducible, strong promoter by comparing the protein profiles of L. casei grown in the presence of 0.3% bile salt (mimicking intestine conditions). Six spots that were consistently overexpressed in the presence of bile salts were isolated and identified. Their promoter reigns are now under investigation and characterization.
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Zhou, Ting, Roni Shapira, Peter Pauls, Nachman Paster, and Mark Pines. Biological Detoxification of the Mycotoxin Deoxynivalenol (DON) to Improve Safety of Animal Feed and Food. United States Department of Agriculture, July 2010. http://dx.doi.org/10.32747/2010.7613885.bard.

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The trichothecene deoxynivalenol (DON, vomitoxin), one of the most common mycotoxin contaminants of grains, is produced by members of the Fusarium genus. DON poses a health risk to consumers and impairs livestock performance because it causes feed refusal, nausea, vomiting, diarrhea, hemolytic effects and cellular injury. The occurrence of trichothecenes contamination is global and they are very resistant to physical or chemical detoxification techniques. Trichothecenes are absorbed in the small intestine into the blood stream. The overall objective of this project was to develop a protecting system using probiotic bacteria that will express trichothecene 3-O-acetyltransferase (Tri101) that convert T-2 to a less toxic intermediate to reduce ingested levels in-situ. The major obstacle that we had faced during the project is the absence of stable and efficient expression vectors in probiotics. Most of the project period was invested to screen and isolate strong promoter to express high amounts of the detoxify enzyme on one hand and to stabilize the expression vector on the other hand. In order to estimate the detoxification capacity of the isolated promoters we had developed two very sensitive bioassays.The first system was based on Saccharomyces cerevisiae cells expressing the green fluorescent protein (GFP). Human liver cells proliferation was used as the second bioassay system.Using both systems we were able to prove actual detoxification on living cells by probiotic bacteria expressing Tri101. The first step was the isolation of already discovered strong promoters from lactic acid bacteria, cloning them downstream the Tri101 gene and transformed vectors to E. coli, a lactic acid bacteria strain Lactococcuslactis MG1363, and a probiotic strain of Lactobacillus casei. All plasmid constructs transformed to L. casei were unstable. The promoter designated lacA found to be the most efficient in reducing T-2 from the growth media of E. coli and L. lactis. A prompter library was generated from L. casei in order to isolate authentic probiotic promoters. Seven promoters were isolated, cloned downstream Tri101, transformed to bacteria and their detoxification capability was compared. One of those prompters, designated P201 showed a relatively high efficiency in detoxification. Sequence analysis of the promoter region of P201 and another promoter, P41, revealed the consensus region recognized by the sigma factor. We further attempted to isolate an inducible, strong promoter by comparing the protein profiles of L. casei grown in the presence of 0.3% bile salt (mimicking intestine conditions). Six spots that were consistently overexpressed in the presence of bile salts were isolated and identified. Their promoter reigns are now under investigation and characterization.
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Avni, Adi, and Gitta L. Coaker. Proteomic investigation of a tomato receptor like protein recognizing fungal pathogens. United States Department of Agriculture, January 2015. http://dx.doi.org/10.32747/2015.7600030.bard.

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Maximizing food production with minimal negative effects on the environment remains a long-term challenge for sustainable food production. Microbial pathogens cause devastating diseases, minimizing crop losses by controlling plant diseases can contribute significantly to this goal. All plants possess an innate immune system that is activated after recognition of microbial-derived molecules. The fungal protein Eix induces defense responses in tomato and tobacco. Plants recognize Eix through a leucine-rich-repeat receptor- like-protein (LRR-RLP) termed LeEix. Despite the knowledge obtained from studies on tomato, relatively little is known about signaling initiated by RLP-type immune receptors. The focus of this grant proposal is to generate a foundational understanding of how the tomato xylanase receptor LeEix2 signals to confer defense responses. LeEix2 recognition results in pattern triggered immunity (PTI). The grant has two main aims: (1) Isolate the LeEix2 protein complex in an active and resting state; (2) Examine the biological function of the identified proteins in relation to LeEix2 signaling upon perception of the xylanase elicitor Eix. We used two separate approaches to isolate receptor interacting proteins. Transgenic tomato plants expressing LeEix2 fused to the GFP tag were used to identify complex components at a resting and activated state. LeEix2 complexes were purified by mass spectrometry and associated proteins identified by mass spectrometry. We identified novel proteins that interact with LeEix receptor by proteomics analysis. We identified two dynamin related proteins (DRPs), a coiled coil – nucleotide binding site leucine rich repeat (SlNRC4a) protein. In the second approach we used the split ubiquitin yeast two hybrid (Y2H) screen system to identified receptor-like protein kinase At5g24010-like (SlRLK-like) (Solyc01g094920.2.1) as an interactor of LeEIX2. We examined the role of SlNRC4a in plant immunity. Co-immunoprecipitation demonstrates that SlNRC4a is able to associate with different PRRs. Physiological assays with specific elicitors revealed that SlNRC4a generally alters PRR-mediated responses. SlNRC4a overexpression enhances defense responses while silencing SlNRC4 reduces plant immunity. We propose that SlNRC4a acts as a non-canonical positive regulator of immunity mediated by diverse PRRs. Thus, SlNRC4a could link both intracellular and extracellular immune perception. SlDRP2A localizes at the plasma membrane. Overexpression of SlDRP2A increases the sub-population of LeEIX2 inVHAa1 endosomes, and enhances LeEIX2- and FLS2-mediated defense. The effect of SlDRP2A on induction of plant immunity highlights the importance of endomembrane components and endocytosis in signal propagation during plant immune . The interaction of LeEIX2 with SlRLK-like was verified using co- immunoprecipitation and a bimolecular fluorescence complementation assay. The defence responses induced by EIX were markedly reduced when SlRLK-like was over-expressed, and mutation of slrlk-likeusing CRISPR/Cas9 increased EIX- induced ethylene production and SlACSgene expression in tomato. Co-expression of SlRLK-like with different RLPs and RLKs led to their degradation, apparently through an endoplasmic reticulum-associated degradation process. We provided new knowledge and expertise relevant to expression of specific be exploited to enhance immunity in crops enabling the development of novel environmentally friendly disease control strategies.
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Eyal, Yoram, and Sheila McCormick. Molecular Mechanisms of Pollen-Pistil Interactions in Interspecific Crossing Barriers in the Tomato Family. United States Department of Agriculture, May 2000. http://dx.doi.org/10.32747/2000.7573076.bard.

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During the evolutionary process of speciation in plants, naturally occurring barriers to reproduction have developed that affect the transfer of genes within and between related species. These barriers can occur at several different levels beginning with pollination-barriers and ending with hybrid-breakdown. The interaction between pollen and pistils presents one of the major barriers to intra- and inter-specific crosses and is the focus of this research project. Our long-term goal in this research proposal was defined to resolve questions on recognition and communication during pollen-pistil interactions in the extended tomato family. In this context, this work was initiated and planned to study the potential involvement of tomato pollen-specific receptor-like kinases (RLK's) in the interaction between pollen and pistils. By special permission from BARD the objectives of this research were extended to include studies on pollen-pistil interactions and pollination barriers in horticultural crops with an emphasis on citrus. Functional characterization of 2 pollen-specific RLK's from tomato was carried out. The data shows that both encode functional kinases that were active as recombinant proteins. One of the kinases was shown to accumulate mainly after pollen germination and to be phosphorylated in-vitro in pollen membranes as well as in-vivo. The presence of style extract resulted in dephosphorylation of the RLK, although no species specificity was observed. This data implies a role for at least one RLK in pollination events following pollen germination. However, a transgenic plant analysis of the RLK's comprising overexpression, dominant-negative and anti-sense constructs failed to provide answers on their role in pollination. While genetic effects on some of the plants were observed in both the Israeli and American labs, no clear functional answers were obtained. An alternative approach to addressing function was pursued by screening for an artificial ligand for the receptor domain using a peptide phage display library. An enriched peptide sequence was obtained and will be used to design a peptide-ligand to be tested for its effect o pollen germination and tube growth. Self-incompatibility (SI) in citrus was studied on 3 varieties of pummelo. SI was observed using fluorescence microscopy in each of the 3 varieties and compatibility relations between varieties was determined. An initial screen for an S-RNase SI mechanism yielded only a cDNA homologous to the group of S-like RNases, suggesting that SI results from an as yet unknown mechanism. 2D gel electrophoresis was applied to compare pollen and style profiles of different compatibility groups. A "polymorphic" protein band from style extracts was observed, isolated and micro-sequenced. Degenerate primers designed based on the peptide sequence date will be used to isolate the relevant genes i order to study their potential involvement in SI. A study on SI in the apple cultivar Top red was initiated. SI was found, as previously shown, to be complete thus requiring a compatible pollinator variety. A new S-RNase allele was discovered fro Top red styles and was found to be highly homologous to pear S-RNases, suggesting that evolution of these genes pre-dated speciation into apples and pears but not to other Rosaceae species. The new allele provides molecular-genetic tools to determine potential pollinators for the variety Top red as well as a tool to break-down SI in this important variety.
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Litaor, Iggy, James Ippolito, Iris Zohar, and Michael Massey. Phosphorus capture recycling and utilization for sustainable agriculture using Al/organic composite water treatment residuals. United States Department of Agriculture, January 2015. http://dx.doi.org/10.32747/2015.7600037.bard.

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Objectives: 1) develop a thorough understanding of the sorption mechanisms of Pi and Po onto the Al/O- WTR; 2) determine the breakthrough range of the composite Al/O-WTR during P capturing from agro- wastewaters; and 3) critically evaluate the performance of the composite Al/O-WTR as a fertilizer using selected plants grown in lysimeters and test-field studies. Instead of lysimeters we used pots (Israel) and one- liter cone-tainers (USA). We conducted one field study but in spite of major pretreatments the soils still exhibited high enough P from previous experiments so no differences between control and P additions were noticeable. Due to time constrains the field study was discontinued. Background: Phosphorous, a non-renewable resource, has been applied extensively in fields to increase crop yield, yet consequently has increased the potential of waterway eutrophication. Our proposal impetus is the need to develop an innovative method of P capturing, recycling and reuse that will sustain agricultural productivity while concurrently reducing the level of P discharge from and to agricultural settings. Major Conclusions & Achievements: An innovative approach was developed for P removal from soil leachate, dairy wastewater (Israel), and swine effluents (USA) using Al-based water treatment residuals (Al- WTR) to create an organic-Al-WTR composite (Al/O-WTR), potentially capable of serving as a P fertilizer source. The Al-WTR removed 95% inorganic-P, 80% to 99.9% organic P, and over 60% dissolved organic carbon from the agro-industrial waste streams. Organic C accumulation on particles surfaces possibly enhanced weak P bonding and facilitated P desorption. Analysis by scanning electron microscope (SEM- EDS), indicated that P was sparsely sorbed on both calcic and Al (hydr)oxide surfaces. Sorption of P onto WW-Al/O-WTR was reversible due to weak Ca-P and Al-P bonds induced by the slight alkaline nature and in the presence of organic moieties. Synchrotron-based microfocused X-ray fluorescence (micro-XRF) spectrometry, bulk P K-edge X-ray absorption near edge structure spectroscopy (XANES), and P K-edge micro-XANES spectroscopy indicated that adsorption was the primary P retention mechanism in the Al- WTR materials. However, distinct apatite- or octocalciumphosphatelike P grains were also observed. Synchrotron micro-XRF mapping further suggested that exposure of the aggregate exteriors to wastewater caused P to diffuse into the porous Al-WTR aggregates. Organic P species were not explicitly identified via P K-edge XANES despite high organic matter content, suggesting that organic P may have been predominantly associated with mineral surfaces. In screen houses experiments (Israel) we showed that the highest additions of Al/O-WTR (5 and 7 g kg⁻¹) produced the highest lettuce (Lactuca sativa L. var. longifolial) yield. Lettuce yield and P concentration were similar across treatments, indicating that Al/O- WTR can provide sufficient P to perform similarly to common fertilizers. A greenhouse study (USA) was utilized to compare increasing rates of swine wastewater derived Al/O-WTR and inorganic P fertilizer (both applied at 33.6, 67.3, and 134.5 kg P₂O₅ ha⁻¹) to supply plant-available P to spring wheat (TriticumaestivumL.) in either sandy loam or sandy clay loam soil. Spring wheat straw and grain P uptake were comparable across all treatments in the sandy loam, while Al/O-WTR application to the sandy clay loam reduced straw and grain P uptake. The Al/O-WTR did not affect soil organic P concentrations, but did increase phosphatase activity in both soils; this suggests that Al/O-WTR application stimulated microorganisms and enhance the extent to which microbial communities can mineralize Al/O-WTR-bound organic P. Implications: Overall, results suggest that creating a new P fertilizer from Al-WTR and agro-industrial waste sources may be a feasible alternative to mining inorganic P fertilizer sources, while protecting the environment from unnecessary waste disposal.
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Weinberg, Zwi G., Adegbola Adesogan, Itzhak Mizrahi, Shlomo Sela, Kwnag Jeong, and Diwakar Vyas. effect of selected lactic acid bacteria on the microbial composition and on the survival of pathogens in the rumen in context with their probiotic effects on ruminants. United States Department of Agriculture, January 2014. http://dx.doi.org/10.32747/2014.7598162.bard.

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This research project was performed in context of the apparent probiotic effect of selected lactic acid bacteria (LAB) silage inoculants on the performance of ruminants (improved feed intake, faster live-weight gain, higher milk yields and improved feed efficiency). The overall objective was to find out how LAB affect ruminant performance. The project included several “chapters” as follows: 1. The effect of LAB silage inoculants on the survival of detrimental bacteria in rumen fluid, in vitro study (Weinberg et al., The Volcani Center). An in vitro model was developed to study the interaction between selected LAB and an E. coli strain tagged with green fluorescence protein (GFP) in buffered RF. Results indicated that both LAB inoculants and E. coli survived in the RF for several days; both LAB inoculants and LAB-treated silages did not affect survival of E. coli in rumen fluid in vitro. The effect of feeding baled wheat silages treated with or without three selected LAB silage inoculants on the performance of high-lactating cows (Weinberg et al., The Volcani Center). Treatments included control (no additive), Lacobacillusbuchneri40788 (LB), Lactobacillus plantarumMTD1 40027 (LP) and Pediococcuspentosaceus30168 (PP), each applied at 10⁶ cfu/g FM. The silages were included in the TMR of 32 high milking Holstein cows in a controlled feeding experiment. All baled silages were of good quality. The LB silage had the numerically highest acetic acid and were the most stable upon aerobic exposure. The cows fed the LB silages had the highest daily milk yields, percent milk fat and protein. The microbiome of baled wheat silages and changes during ensiling of wheat and corn (Sela et al., The Volcani Center). Bacterial community of the baled silages was dominated mainly of two genera in total, dominated by Lactobacillus and Clostridium_sensu_stricto_12 with 300 other genera at very low abundance. Fungal community was composed mainly of two genera in total, dominated by Candida and Monascuswith 20 other genera at very low abundance. In addition, changes in the microbiome during ensiling of wheat and corn with and without addition of L. plantarumMTD1 was studied in mini-silos. Overall 236 bacterial genera were identified in the fresh corn but after 3 months Lactobacillus outnumbered all other species by acquiring 95% of relative abundance. The wheat silage samples are still under analysis. The effect of applying LAB inoculants at ensiling on survival of E. coli O157:H7 in alfalfa and corn silages(Adesogan et al., University of Florida). E. coli (10⁵ cfu/g) was applied to fresh alfalfa and corn at ensiling with or without L. plantarumor L. buchneri. The pathogen was added again after about 3 moths at the beginning of an aerobic exposure period. The inoculants resulted in faster decrease in pH as compared with the control (no additives) or E. coli alone and therefore, the pathogen was eliminated faster from these silages. After aerobic exposure the pathogen was not detected in the LAB treated silages, whereas it was still present in the E. coli alone samples. 5. The effect of feeding corn silage treated with or without L. buchnerion shedding of E. coli O157:H7 by dairy cows (Adesogan et al., UFL). BARD Report - Project 4704 Page 2 of 12 Five hundred cows from the dairy herd of the University of Florida were screened for E. coli shedding, out of which 14 low and 13 high shedders were selected. These cows were fed a total mixed ration (TMR) which was inoculated with E. coli O157:H7 for 21 days. The TMR included corn silage treated with or without L. buchneri. The inoculated silages were more stable upon aerobic exposure than the control silages; the silage inoculant had no significant effect on any milk or cow blood parameters. However, the silage inoculant tended to reduce shedding of E. coli regardless of high or low shedders (p = 0.06). 6. The effect of feeding baled wheat silages treated with or without three selected LAB silage inoculants on the rumen microbiome (Mizrahi et al., BGU). Rumen fluid was sampled throughout the feeding experiment in which inoculated wheat silages were included in the rations. Microbial DNA was subsequently purified from each sample and the 16S rRNA was sequenced, thus obtaining an overview of the microbiome and its dynamic changes for each experimental treatment. We observed an increase in OTU richness in the group which received the baled silage inoculated with Lactobacillus Plantarum(LP). In contrast the group fed Lactobacillus buchneri(LB) inoculated silage resulted in a significant decrease in richness. Lower OTU richness was recently associated in lactating cows with higher performance (Ben Shabatet al., 2016). No significant clustering could be observed between the different inoculation treatments and the control in non metric multi-dimentional scaling, suggesting that the effect of the treatments is not the result of an overall modulation of the microbiome composition but possibly the result of more discrete interactions. Significant phylum level changes in composition also indicates that no broad changes in taxa identity and composition occurred under any treatment A more discrete modulation could be observed in the fold change of several taxonomic groups (genus level analysis), unique to each treatment, before and after the treatment. Of particular interest is the LB treated group, in which several taxa significantly decreased in abundance. BARD Report - Project 4704 Page 3 of 12
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